Cell monolayers infected with Ad-CMV-hChR2(H134R)-eYFP (MOIs 25 and 250, respectively) or AAV9.CAG.hChR2(H134R)-mCherry.WPRE.SV40 (MOI 50,000–100,000 ± 500 mU/mL NM) were stained with the calcium- and voltage-sensitive dyes and optically mapped using our recently published all-optical, high-throughput system for dynamic cardiac electrophysiology, termed OptoDyCE (Klimas et al., 2016 (link), 2018 (link)). The excitation filter for the actuating LED was 470/28 nm, the LED illumination for the voltage (di-4-ANBDQBS or Berst1) and calcium (Rhod-4AM) measurements was filtered as follows: 655/40 nm and 535/50, respectively. Fluorescence was collected by iXon Ultra 897 EMCCD; Andor, after passing through the emission filter 595/40 nm+700LP. Note that the UPenn Core considers the CAG and the CB7 promoters equivalent and uses them interchangeably; both are ubiquitous promoters, derivatives of CMV (Miyazaki et al., 1989 (link)).
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